PHAs are bacterial polyesters that accumulate in a wide variety of bacteria. These polymers have properties ranging from stiff and brittle plastics to rubber-like materials, and are biodegradable. Because of these properties, PHAs are an attractive source of nonpolluting plastics and elastomers.
The present invention especially relates to the production of copolyesters of .beta.-hydroxybutyrate (3HB) and .beta.-hydroxyvalerate (3HV), designated P(3HB-co-3HV) copolymer, and derivatives thereof.
Currently, there are approximately a dozen biodegradable plastics in commercial use that possess properties suitable for producing a number of specialty and commodity products (Lindsay, 1992). One such biodegradable plastic in the polyhydroxyalkanoate (PHA) family that is commercially important is Biopol.TM., a random copolymer of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). This bioplastic is used to produce biodegradable molded material (e.g., bottles), films, coatings, and in drug release applications. Biopol.TM. is produced via a fermentation process employing the bacterium Alcaigenes eutrophus (Byrom, 1987). The current market price is $6-7/lb, and the annual production is 1,000 tons. By best estimates, this price can be reduced only about 2-fold via fermentation (Poirier et al., 1995). Competitive synthetic plastics such as polypropylene and polyethylene cost about 35-45.cent./lb (Layman, 1994). The annual global demand for polyethylene alone is about 37 million metric tons (Poirier et al., 1995). It is therefore likely that the cost of producing P(3HB-co-3HV) by microbial fermentation will restrict its use to low-volume specialty applications.
Nakamura et al. (1992) reported using threonine (20 g/L) as the sole carbon source for the production of P(3HB-co-3HV) copolymer in A. eutrophus. These workers initially suggested that the copolymer might form via the degradation of threonine by threonine deaminase, with conversion of the resultant .alpha.-ketobutyrate to propionyl-CoA. However, they ultimately concluded that threonine was utilized directly, without breaking carbon-carbon bonds, to form valeryl-CoA as the 3HV precursor. The nature of this chemical conversion was not described, but since the breaking of carbon-carbon bonds was not postulated to occur, the pathway could not involve threonine deaminase in conjunction with an .alpha.-ketoacid decarboxylating step to form propionate or propionyl-CoA. In the experiments of Nakamura et al, the PHA polymer content was very low (&lt;6% of dry cell weight). This result, in conjunction with the expense of feeding bacteria threonine, makes their approach impractical for the commercial production of P(3HB-co3HV) copolymer.
Yoon et al. (1995) have shown that growth of Alcaligenes sp. SH-69 on a medium supplemented with threonine, isoleucine, or valine resulted in significant increases in the 3HV fraction of the P(3HB-co-3HV) copolymer. In addition to these amino acids, glucose (3% wt/vol) was also added to the growth media. In contrast to the results obtained by Nakamura et al. (1992), growth of A. eutrophus under the conditions described by Yoon et al. (1995) did not result in the production of P(3HB-co-3HV) copolymer when the medium was supplemented with threonine as the sole carbon source. From their results, Yoon et al. (1995) implied that the synthetic pathway for the 3HV component in P(3HB-co-3HV) copolymer is likely the same as that described in WO 91/18995 and Steinbuchel and Pieper (1992). This postulated synthetic pathway involves the degradation of isoleucine to propionyl-CoA (FIG. 3).
U.S. Pat. No. 5,602,321 teaches the insertion and expression of polymer biosynthesis genes in plants, and preferably in cotton. Genetic constructs encoding ketothiolase, acetoacetyl CoA reductase, and PHB synthase enzymes were introduced into cotton.
Sim et al. (1997) reported that the amount of polyhydroxyalkanoate synthase protein in a bacterial cell affected the molecular weight and polydispersity of polymers produced therein. Increased synthase concentrations led to the biosynthesis of lower molecular weight polymer.
John and Keller (1996) obtained polyhydroxybutyrate from cotton transformed with the phaB and phaC genes from Alcaligenes eutrophus. A major fraction of the polymer obtained had a molecular weight in excess of 600,000. Polydispersity of the polymer is not discussed, nor is sufficient data presented to allow calculation thereof.
Poirier et al. (1995b) described the biosynthesis of polyhydroxybutyrate in a suspension culture of Arabidopsis thaliana plant cells expressing the phbB and phbC genes from Alcaligenes eutrophus. No C3 or C5 3-hydroxyacids were detected by gas chromatography of plant extracts. The polyhydroxybutyrate was found to have a broad molecular weight distribution of 10.5. Unlike bacterially produced polymer, the plant cell produced material displayed a polymodal distribution, comprising at least three distinct subpopulations of molecular weight ranges.
The PHB Biosynthetic Pathway
Polyhydroxybutyrate (PHB) was first discovered in 1926 as a constituent of the bacterium Bacillus megaterium (Lemoigne, 1926). Since then, PHAs such as PHB have been found in more than 90 different genera of gram-negative and gram-positive bacteria (Steinbuchel, 1991). These microorganisms produce PHAs using R-.beta.-hydroxyacyl-CoAs as the direct metabolic substrate for a PHA synthase, and produce polymers of R-(3)-bydroxyalkanoates having chain lengths ranging from C3-C14 (Steinbuchel and Valentin, 1995).
To date, the best understood biochemical pathway for PHB production is that found in the bacterium Alcaligenes eutrophus (Dawes and Senior, 1973; Slater et al., 1988; Schubert et al., 1988; Peoples and Sinskey, 1989a and 1989b). This pathway, which is also utilized by other microorganisms, is summarized in FIG. 1. In this organism, an operon encoding three gene products, i.e., PHB synthase, .beta.-ketothiolase, and acetoacetyl-CoA reductase, encoded by the phbC, phbA, and phbB genes, respectively, are required to produce the PHA homopolymer R-polyhydroxybutyrate (PHB).
As further shown in FIG. 1, acetyl-CoA is the starting substrate employed in the biosynthetic pathway. This metabolite is naturally available for PHB production in plants and bacteria when these organisms are genetically manipulated to produce the PHB polyester.
Recently, a multi-enzyme pathway was successfully introduced into plants for the generation of polyhydroxybutyrate (PHB) (Poirier et al., 1992).
The P(3HB-co-3HV) Copolymer Biosynthetic Pathway
As noted above, P(3HB-co-3HV) random copolymer, commercially known as Biopol.TM., is produced by fermentation employing A. eutrophus. A proposed biosynthetic pathway for P(3HB-co-3HV) copolymer production is shown in FIG. 2. Production of this polymer in plants has not yet been demonstrated.
The successful production of P(3HB-co-3HV) copolymer in plants or bacteria requires the generation of substrates that can be utilized by the PHA biosynthetic enzymes. For the 3HB component of the polymer, the substrate naturally exists in plants in sufficient concentration in the form of acetyl-CoA (Nawrath et al., 1994). This is not true for the 3HV component of the copolymer, however. In this case, the starting substrate is propionyl-CoA. The presence of sufficient pools of acetyl-CoA and propionyl-CoA in plants and microorganisms, along with the proper PHA biosynthetic enzymes (i.e., a .beta.-ketothiolase, a .beta.-ketoacyl-CoA reductase, and a PHA synthase), would make it possible to produce copolyesters of P(3HB-co-3HV) in these organisms.
The present invention provides a variety of different methods for optimizing levels of substrates employed in the biosynthesis of copolymers of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) in plants and bacteria via manipulation of normal metabolic pathways using recombinant DNA techniques.
In one aspect, the present invention provides methods for the production of enhanced levels of threonine, .alpha.-ketobutyrate, propionyl-CoA, .beta.-ketovaleryl-CoA, and .beta.-hydroxyvaleryl-CoA, all of which are metabolites in the biosynthetic pathway of 3HV in the P(3HB-co-3HV) copolymer, from various carbon sources in plants or bacteria. This is achieved by providing a variety of wild-type and/or deregulated enzymes involved in the biosynthesis of the aspartate family of amino acids (i.e., aspartate, threonine, lysine, and methionine), and wild-type or deregulated forms of enzymes involved in the conversion of threonine to P(3HB-co-3HV) copolymer endproduct. Using these enzymes, the levels of the above-noted metabolites can be increased in plants and bacteria in the range of from about 1-10 fold, 1-100 fold, or 1-1000 fold.
In another aspect, the present invention provides methods for the biological production of P(3HB-co-3HV) copolymers in plants and bacteria utilizing propionyl-CoA as a substrate. As shown in FIG. 3, propionyl-CoA can be produced through a variety of engineered metabolic pathways. Introduction into plants and bacteria of appropriate .beta.-ketothiolases capable of condensing acetyl-CoA with itself and/or with propionyl-CoA, along with appropriate .beta.-ketoacyl-CoA reductases and PHA synthases, in combination with various enzymes involved in asparate family amino acid biosynthesis and the conversion of threonine to PHA copolymer precursors, will permit these organisms to produce P(3HB-co-3HV) copolymers. The PHA biosynthetic enzymes can be those of A. eutrophus or other organisms, or enzymes that catalyze reactions involved in fatty acid biosynthesis or degradation, ultimately resulting in the conversion of acetyl-CoA and propionyl-CoA to P(3HB-co-3HV). In plants, these enzymes can be expressed in the cytoplasm or targeted to organelles such as plastids (e.g., those of leaves or seeds) or mitochondria via the use of transit peptides for enhanced production of polyhydroxyalkanoates. Alternatively, plastids can be transformed with recombinant constructs that facilitate expression of these enzymes directly within the plastids themselves.
The enzymes discussed herein can be employed alone or in various combinations in order to enhance the levels of threonine, .alpha.-ketobutyrate, propionate, propionyl-CoA, .beta.-ketovaleryl-CoA, and .beta.-hydroxyvaleryl-CoA, and for the production of P(3HB-co-3HV) copolymer.
More specifically, the present invention encompasses the following aspects:
An isoleucine-deregulated mutein of E. coli threonine deaminase, wherein leucine at amino acid position 447 is replaced with an amino acid selected from the group consisting of alanine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine. In one aspect, leucine at amino acid position 447 can be replaced with phenylalanine.
An isoleucine-deregulated mutein of E. coli threonine deaminase, wherein leucine at amino acid position 447 is replaced with an amino acid selected from the group consisting of alanine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine, and leucine at amino acid position 481 is replaced with an amino acid selected from the group consisting of alanine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine. In one aspect, leucine at amino acid position 447 can be replaced with phenylalanine, and leucine at amino acid position 481 can be replaced with phenylalanine.
An .alpha.-ketoacid dehydrogenase complex, comprising an .alpha.-ketoacid decarboxylase E1 subunit, a dihydrolipoyl transacylase E2 subunit, and a dihydrolipoyl dehydrogenase E3 subunit, wherein said .alpha.-ketoacid decarboxylase E1 subunit exhibits improved binding and decarboxylating properties with .alpha.-ketobutyrate compared to pyruvate decarboxylase E1 subunit naturally present in a host cell pyruvate dehydrogenase complex. The .alpha.-ketoacid decarboxylase E1 subunit can be a branched-chain .alpha.-ketoacid decarboxylase E1 subunit such as bovine kidney, Pseudomonas putida, or Bacillus subtilis branched-chain .alpha.-ketoacid dehydrogenase E1 subunit.
An .alpha.-ketoacid dehydrogenase complex, comprising an .alpha.-ketoacid decarboxylase E1 subunit, a dihydrolipoyl transacylase E2 subunit, and a dihydrolipoyl dehydrogenase E3 subunit, wherein said E1 and E2 subunits exhibit improved binding/decarboxylating and transacylase properties, respectively, with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit and dihydrolipoyl transacetylase E2 subunit, respectively, naturally present in a host cell pyruvate dehydrogenase complex. The .alpha.-ketoacid decarboxylase E1 and dihydrolipoyl transacylase E2 subunits can be branched-chain .alpha.-ketoacid dehydrogenase E1 and E2 subunits such as those from bovine kidney, Pseudomonas putida, or Bacillus subtilis.
An isolated .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA.
An isolated .beta.-ketothiolase capable of condensing acetyl-CoA and butyryl-CoA to produce .beta.-ketocaproyl-CoA.
An isolated .beta.-ketothiolase capable of:
condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA; PA1 condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; and PA1 condensing acetyl-CoA and butyryl-CoA to produce .beta.-ketcaproyl-CoA. PA1 (a) the nucleotide sequence shown in SEQ ID NO:9 or the complement thereof; PA1 (b) a nucleotide sequence that hybridizes to said nucleotide sequence of (a) under a wash stringency equivalent to 0.5.times.SSC to 2.times.SSC, 0.1% SDS, at 55-65.degree. C., and which encodes an enzyme having enzymatic activity similar to that of A. eutrophus BktB .beta.-ketothiolase; PA1 (c) a nucleotide sequence encoding the same genetic information as said nucleotide sequence of (a), but which is degenerate in accordance with the degeneracy of the genetic code; and PA1 (d) a nucleotide sequence encoding the same genetic information as said nucleotide sequence of (b), but which is degenerate in accordance with the degeneracy of the genetic code. PA1 wherein said introduced DNA is operatively linked to regulatory signals that cause expression of said introduced DNA; and PA1 wherein cells of said plant produce an elevated amount of threonine compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNA. PA1 a wild-type or deregulated aspartate kinase enzyme; and PA1 threonine synthase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of threonine compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; and PA1 homoserine dehydrogenase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of threonine compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; and PA1 threonine synthase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of threonine compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of .alpha.-ketobutyrate compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 threonine synthase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of .alpha.-ketobutyrate compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of .alpha.-keto-butyrate compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of .alpha.-keto-butyrate compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit enzyme that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex of said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; and PA1 a dihydrolipoyl transacylase E2 subunit exhibiting improved transacylase activity with .alpha.-ketobutyrate compared with dihydrolipoyl transacetylase E2 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; and PA1 pyruvate oxidase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of propionyl-CoA compared that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 pyruvate oxidase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an acyl-CoA synthetase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an acyl-CoA synthetase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an elevated amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said plant produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said plant produces P(3HB-co-3HV) copolymer. PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said plant produces P(3HB-co-3HV) copolymer. PA1 wherein said introduced DNA is operatively linked to regulatory signals that cause expression of said introduced DNA; and PA1 wherein said plant produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said plant produces P(3HB-co-3HV) copolymer. PA1 wherein said introduced DNA is operatively linked to regulatory signals that cause expression of said introduced DNA; and PA1 wherein cells of said plant produce an increased amount of .alpha.-keto-butyrate compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNA. PA1 threonine synthase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of .alpha.-keto-butyrate compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; and PA1 pyruvate oxidase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 pyruvate oxidase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an acyl-CoA synthetase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an acyl-CoA synthetase; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein cells of said plant produce an increased amount of propionyl-Co A compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-Co A compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce an increased amount of propionyl-CoA compared to that in cells of a corresponding, wild-type plant not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said plant; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein cells of said plant produce P(3HB-co-3HV) copolymer. PA1 wherein each of said introduced DNAs is operatively linked to a plastid transit peptide coding region capable of directing transport of said enzymes into a plastid, and regulatory signals that cause expression of said introduced DNAs in seeds of said plant; and PA1 wherein P(3HB) homopolymer is produced in seeds of said plant. PA1 wherein said introduced DNA is operatively linked to regulatory signals that cause expression of said introduced DNA; and PA1 wherein said bacterium produces an elevated amount of threonine compared to that in a corresponding wild-type bacterium not comprising said introduced DNA. PA1 a wild-type or deregulated aspartate kinase; and PA1 threonine synthase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of threonine compared to that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; and PA1 homoserine dehydrogenase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of threonine compared to that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; and PA1 threonine synthase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of threonine compared to that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of .alpha.-keto-butyrate compared to that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 threonine synthase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of .alpha.-keto-butyrate compared to that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of .alpha.-keto-butyrate compared to that in a wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of .alpha.-keto-butyrate compared to that in a wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex of said bacterium; PA1 wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of propionyl-CoA compared to that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 a pyruvate decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; and PA1 a dihydrolipoyl transacylase E2 subunit exhibiting improved transacylase activity with .alpha.-ketobutyrate compared with dihydrolipoyl transacetylase E2 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of propionyl-CoA compared to that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; and PA1 pyruvate oxidase; PA1 wherein said introduced DNAs are operably linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of propionyl-CoA compared that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 pyruvate oxidase; PA1 wherein said introduced DNAs are operably linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an elevated amount of propionyl-CoA compared that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 wherein said introduced DNA is operatively linked to regulatory signals that cause expression of said introduced DNA; and PA1 wherein said bacterium produces an increased amount of .alpha.-keto-butyrate compared to that in a corresponding wild-type bacterium not comprising said introduced DNA. PA1 threonine synthase; and PA1 a wild-type or deregulated threonine deaminase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of .alpha.-keto-butyrate compared to that in a corresponding wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; and PA1 pyruvate oxidase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 pyruvate oxidase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an acyl-CoA synthetase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an acyl-CoA synthetase; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-Co A compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-Co A compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in is a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; and PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces an increased amount of propionyl-CoA compared to that in a corresponding, wild-type bacterium not comprising said introduced DNAs. PA1 a wild-type or deregulated aspartate kinase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 a wild-type or deregulated aspartate kinase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type, partially, or totally lysine feedback inhibition insensitive aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type, partially, or totally lysine feedback inhibition insensitive aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating, activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a wild-type or deregulated aspartate kinase; PA1 homoserine dehydrogenase; PA1 threonine synthase; PA1 a wild-type or deregulated threonine deaminase; PA1 pyruvate oxidase; PA1 an acyl-CoA synthetase; PA1 an .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit naturally present in pyruvate dehydrogenase complex in said bacterium; PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer. PA1 culturing any of the four foregoing mentioned bacteria under conditions and for a time conducive to the formation of P(3HB-co-3HV) copolymer; and PA1 recovering said P(3HB-co-3HV) copolymer produced thereby. PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA and a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; or PA1 a .beta.-ketothiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-CoA, and capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA; PA1 a .beta.-ketoacyl-CoA reductase capable of reducing acetoacetyl-CoA and .beta.-ketovaleryl-CoA to produce .beta.-hydroxybutyryl-CoA and .beta.-hydroxy-valeryl-CoA, respectively; and PA1 a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer; PA1 wherein said introduced DNAs are operatively linked to regulatory signals that cause expression of said introduced DNAs; and PA1 wherein said bacterium produces P(3HB-co-3HV) copolymer when grown on a medium comprising propionic acid. PA1 culturing the foregoing bacterium on a medium comprising propionic acid under conditions and for a time conducive to the formation of P(3HB-co-3HV) copolymer, and PA1 recovering said P(3HB-co-3HV) copolymer produced thereby. PA1 (a) preparing a stem explant from a canola plant by: PA1 (b) inserting DNA to be introduced into said explant of step (a) by inoculating said explant with a disarmed Agrobacterium tumefaciens vector containing said DNA; PA1 (c) culturing said explant of step (b) in the basal-side down orientation; PA1 (d) selecting transformed explant tissue; and PA1 (e) regenerating a differentiated transformed plant from said transformed explant tissue of step (d). PA1 (a) preparing a cotyledon explant from a soybean seedling by: PA1 (b) inserting DNA to be introduced into said explant of step (a) by inoculating at least the region adjacent to the axillary bud of the explant with a disarmed Agrobacterium tumefaciens vector containing said DNA; PA1 (c) selecting transformed explant tissue; and PA1 (d) regenerating a differentiated transformed plant from said transformed explant tissue of step (c). PA1 wherein said associated hypocotyl segment is completely separated from its adjacent hypocotyl segment attached to the remaining cotyledon, thus separating said cotyledons; PA1 wherein said epicotyl has been removed from the cotyledon to which it is attached; PA1 wherein the cotyledon is wounded in the region of said axillary bud; and PA1 wherein said seedling has been incubated at a temperature of from about 0.degree. C. to about 10.degree. C. for at least about 24 hours prior to preparing said soybean tissue.
The foregoing isolated .beta.-ketothiolase is exemplified by BktB .beta.-keto-thiolase having the amino acid sequence shown in SEQ ID NO:11.
Isolated DNA molecules comprising a nucleotide sequence encoding the isoleucine-deregulated muteins of E. coli threonine deaminase described herein.
An isolated DNA molecule comprising the nucleotide sequence shown in SEQ ID NO:5.
An isolated DNA molecule comprising the nucleotide sequence shown in SEQ ID NO:8.
An isolated DNA molecule comprising a nucleotide sequence selected from the group consisting of:
The foregoing isolated DNA molecule, wherein said wash stringency is equivalent to 2.times.SSC, 0.1% SDS, at 55.degree. C.
The foregoing isolated DNA molecule, wherein said wash stringency is equivalent to 1.times.SSC, 0.1% SDS, at 55.degree. C.
The foregoing isolated DNA molecule, wherein said wash stringency is equivalent to 0.5.times.SSC, 0.1% SDS, at 55.degree. C.
An isolated DNA molecule, comprising the nucleotide sequence shown in SEQ ID NO:9 or the complement thereof.
A recombinant vector, comprising any of the foregoing nucleotide sequences operatively linked to a promoter and 5' and 3' regulatory sequences sufficient to drive expression of said nucleotide sequences in a host cell.
A recombinant vector, comprising a nucleotide sequence encoding E. coli threonine deaminase wherein leucine at position 481 is replaced with phenylalanine, operatively linked to a promoter and 5' and 3' regulatory sequences sufficient to drive expression of said nucleotide sequence in a host cell.
The foregoing recombinant vector, wherein said nucleotide sequence comprises the sequence shown in SEQ ID NO:7.
A host cell, comprising any of the foregoing recombinant vectors.
A host cell, comprising an .alpha.-ketoacid dehydrogenase complex comprising an .alpha.-ketoacid decarboxylase E1 subunit, a dihydrolipoyl transacylase E2 subunit, and a dihydrolipoyl dehydrogenase E3 subunit, wherein said .alpha.-ketoacid decarboxylase E1 subunit exhibits improved binding and decarboxylating properties with .alpha.-ketobutyrate compared to pyruvate decarboxylase E1 subunit naturally present in a host cell pyruvate dehydrogenase complex, wherein said host cell produces an increased amount of propionyl-CoA compared to a corresponding host cell comprising wild-type pyruvate dehydrogenase complex. The .alpha.-ketoacid decarboxylase E1 subunit which exhibits improved binding and decarboxylating properties with .alpha.-ketobutyrate can be a branched-chain .alpha.-ketoacid decarboxylase E1 subunit.
A host cell, comprising an .alpha.-ketoacid dehydrogenase complex, comprising an .alpha.-ketoacid decarboxylase E1 subunit, a dihydrolipoyl transacylase E2 subunit, and a dihydrolipoyl dehydrogenase E3 subunit, wherein said E1 and E2 subunits exhibit improved binding/decarboxylating and transacylase properties, respectively, with .alpha.-ketobutyrate compared with pyruvate decarboxylase E1 subunit and dihydrolipoyl transacetylase E2 subunit, respectively, naturally present in a host cell pyruvate dehydrogenase complex, wherein said host cell produces an increased amount of propionyl-CoA compared to a corresponding host cell comprising wild-type pyruvate dehydrogenase complex.
A plant, the genome of which comprises introduced DNA encoding a wild-type or deregulated aspartate kinase enzyme;
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
The foregoing plant, wherein said .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate is a branched chain .alpha.-ketoacid decarboxylase E1 subunit such as that from bovine kidney, Pseudomonas putida, or Bacillus subtilis.
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
The foregoing plant, wherein said .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate is a branched chain .alpha.-ketoacid decarboxylase E1 subunit such as that from bovine kidney, Pseudomonas putida, or Bacillus subtilis, and said dihydrolipoyl transacylase E2 subunit exhibiting improved transacylase activity with .alpha.-ketobutyrate is a branched chain dihydrolipoyl transacylase E2 subunit such as that from bovine kidney, Pseudomonas putida, or Bacillus subtilis.
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
The foregoing plant, the genome of which further comprises introduced DNA encoding a wild-type or deregulated threonine deaminase;
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
Any of the foregoing plants, wherein the .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA is BktB.
Any of the foregoing plants, wherein said .beta.-ketoacyl-CoA reductase is obtainable from a microorganism selected from the group consisting of Alcaligenes eutrophus, Alcaligenes faecalis, Aphanothece sp., Azotobacter vinelandii, Bacillus cereus, Bacillus megaterium, Beijerinkia indica, Derxia gummosa, Methylobacterium sp., Microcoleus sp., Nocardia corallina, Pseudomonas cepacia, Pseudomonas extorquens, Pseudomonas oleovorans, Rhodobacter sphaeroides, Rhodobacter capsulatus, Rhodospirillum rubrum, and Thiocapsa pfennigii.
Any of the foregoing plants, wherein said polyhydroxyalkanoate synthase is obtainable from a microorganism selected from the group consisting of Alcaligenes eutrophus, Alcaligenes faecalis, Aphanothece sp., Azotobacter vinelandii, Bacillus cereus, Bacillus megaterium, Beijerinkia indica, Derxia gummosa, Methylobacterium sp., Microcoleus sp., Nocardia corallina, Pseudomonas cepacia, Pseudomonas extorquens, Pseudomonas oleovorans, Rhodobacter sphaeroides, Rhodobacter capsulatus, Rhodospirillum rubrum, and Thiocapsa pfennigii.
A plant, the genome of which comprises introduced DNA encoding a wild-type or deregulated threonine deaminase enzyme;
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
wherein each of said introduced DNAs is operatively linked to regulatory signals that cause expression of said introduced DNAs; and
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
A plant, the genome of which comprises introduced DNAs encoding the following enzymes:
Any of the foregoing plants, wherein each of said introduced DNAs is further operatively linked to a plastid transit peptide coding region capable of directing transport of said enzyme encoded thereby into a plastid.
A method of producing P(3HB-co-3HV)copolymer, comprising growing any of the foregoing plants, the genome of which comprises introduced DNAs encoding a .beta.-ketothiolase, a .beta.-ketoacyl-CoA reductase, and a polyhydroxyalkanoate synthase, and recovering said P(3HB-co-3HV) copolymer produced thereby.
The foregoing method, wherein said P(3HB-co-3HV)copolymer is recovered from leaves or seeds of said plant.
A plant cell containing P(3HB-co-3HV) copolymer.
A plant comprising cells containing P(3HB-co-3HV) copolymer.
Seeds of the foregoing plant.
The foregoing plant, wherein said cells are located in leaves or seeds of said plant.
A plant, the genome of which comprises introduced DNAs encoding the enzymes PhbA, PhbB, and PhbC;
A method of producing P(3HB) homopolymer, comprising growing the foregoing plant, and recovering said P(3HB) homopolymer produced thereby.
The foregoing method, wherein said P(3HB) homopolymer is recovered from seeds of said plant.
A bacterium, the genome of which comprises introduced DNA encoding a wild-type or deregulated aspartate kinase;
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
The foregoing bacterium, wherein said .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate is a branched chain .alpha.-ketoacid decarboxylase E1 subunit such as that from bovine kidney, Pseudomonas putida, or Bacillus subtilis.
A bacterium, the genome of which comprises introduced DNAs encoding:
The foregoing bacterium, wherein said .alpha.-ketoacid decarboxylase E1 subunit that exhibits improved binding and decarboxylating activities with .alpha.-ketobutyrate is a branched chain .alpha.-ketoacid decarboxylase E1 subunit such as that from bovine kidney, Pseudomonas putida, or Bacillus subtilis, and said dihydrolipoyl transacylase E2 subunit exhibiting improved transacylase activity with .alpha.-ketobutyrate is one such as that from bovine kidney, Pseudomonas putida, or Bacillus subtilis.
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
The foregoing bacterium, wherein said pyruvate oxidase is encoded by E. coli poxB.
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
The foregoing bacterium, wherein said .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA is BktB.
The foregoing bacterium, wherein said .beta.-ketoacyl-CoA reductase is acetoacetyl-CoA reductase.
The foregoing bacterium, wherein said polyhydroxyalkanoate synthase is obtainable from a microorganism selected from the group consisting of Alcaligenes eutrophus, Alcaligenes faecalis, Aphanothece sp., Azotobacter vinelandii, Bacillus cereus, Bacillus megaterium, Beijerinkia indica, Derxia gummosa, Methylobacterium sp., Microcoleus sp., Nocardia corallina, Pseudomonas cepacia, Pseudomonas extorquens, Pseudomonas oleovorans, Rhodobacter sphaeroides, Rhodobacter capsulatus, Rhodospirillum rubrum, and Thiocapsa pfennigii.
A bacterium, the genome of which comprises introduced DNA encoding a wild-type or deregulated threonine deaminase;
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
wherein said bacterium produces P(3HB-co-3HV) copolymer.
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A method of producing P(3HB-co-3HV) copolymer, comprising growing any of the foregoing bacteria, the genome of which comprises introduced DNAs encoding a .beta.-ketothiolase, a .beta.-ketoacyl-CoA reductase, and a polyhydroxy-alkanoate synthase, and recovering said P(3HB-co-3HV)copolymer produced thereby.
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
The foregoing bacterium can be one that overproduces threonine. The genome thereof can further comprise, in addition to introduced DNAs encoding the enzymes listed above, introduced DNA encoding a wild-type or deregulated threonine deaminase. Alternatively, the foregoing bacterium can be one that produces propionate or propionyl-CoA at levels useful for producing P(3HB-co-3HV) copolymer.
A method of producing P(3HB-co-3HV) copolymer, comprising:
A bacterium, the genome of which comprises introduced DNAs encoding the following enzymes:
A method of producing P(3HB-co-3HV) copolymer, comprising:
A bacterial cell containing P(3HB-co-3HV) copolymer, wherein said P(3HB-co-3HV) copolymer is produced within said bacterial cell as a result of expression of at least one DNA sequence introduced therein that encodes an enzyme necessary for P(3HB-co-3HV) copolymer synthesis.
The foregoing bacterial cell, wherein said at least one DNA sequence is selected from the group consisting of a DNA sequence encoding a .beta.-keto-thiolase capable of condensing two molecules of acetyl-CoA to produce acetoacetyl-Co-A, a DNA sequence encoding a .beta.-ketothiolase capable of condensing acetyl-CoA and propionyl-CoA to produce .beta.-ketovaleryl-CoA, a DNA sequence encoding a .beta.-ketoacyl-CoA reductase, a DNA sequence encoding a polyhydroxyalkanoate synthase capable of incorporating .beta.-hydroxybutyryl-CoA and .beta.-hydroxyvaleryl-CoA into P(3HB-co-3HV) copolymer, and combinations thereof.
The foregoing bacterial cell, further comprising a DNA sequence introduced therein that encodes a wild-type or deregulated threonine deaminase, wherein said DNA sequence encoding said wild-type or deregulated threonine deaminase is expressed.
A method for transforming canola, comprising:
(i) removing leaves and buds along the stem and removing 4-5 inches of said stem below the flower buds; and PA2 (ii) cutting said 4-5 inches of stem into segments; PA2 (i) incubating said seedling at about 0.degree. C. to about 10.degree. C. for at least 24 hours; PA2 (ii) removing the hypocotyl region by cutting in the region of from about 0.2 to about 1.5 cm below the cotyledonary node; PA2 (ii) splitting and completely separating the remaining attached hypocotyl segment, also thereby separating the two cotyledons; PA2 (iii) removing the epicotyl from the cotyledon to which it remains attached; and PA2 (iv) wounding the cotyledon in the region of said axillary bud;
A method for transforming soybean, comprising:
A soybean explant prepared by steps (a)(i)-(a)(iv) of the preceding method.
Soybean tissue prepared from a seedling cotyledon pair containing an epicotyl, axillary buds, and hypocotyl tissue, comprising a single cotyledon containing an axillary bud and associated hypocotyl segment extending from about 0.2 to about 1.5 cm below the cotyledonary node;
The present invention provides a novel means towards obtaining biosynthetic hydroxyalkanoate polymers. In a preferred embodiment, the polymers have a single mode molecular weight distribution. As used herein, a single mode molecular weight distribution refers to a plot of molecular weight (x-axis) against population (y-axis), resulting in a single peak. Multiple mode molecular weight distributions display two or more peaks in such a plot. The plot is first `smoothed` by generally any mathematically valid model, and preferably by the smoothing function described in "Exploratory Data Analysis" (Tukey, J. W. Addison-Wesley publishers, Reading, Mass., 1977, pp 205-235). The three highest peak heights are selected, and their peak heights added together to form a peak sum. In order for a plot to represent a single mode molecular weight distribution, the highest peak height must comprise at least about 80% of the peak sum. Molecular weight may be determined by methods such as size exclusion chromatography or gel permeation chromatography. In a further preferred embodiment, the polymers have a small molecular weight distribution. As used herein, molecular weight distribution, also referred to as polydispersity, is calculated by dividing the weight average molecular weight by the number average molecular weight.
In a preferred embodiment, a plant extract will contain a polyhydroxyalkanoate polymer wherein the polyhydroxyalkanoate polymer was produced by a plant, and where the polyhydroxyalkanoate polymer has a single mode molecular weight distribution. As used herein, a plant extract refers to materials prior to chromatographic separation. The extract may be a plant lysate, or the materials dissolved from contacting the plant or plant materials with a suitable solvent including, but not limited to alcohol or chloroform. Lysates may be prepared by a variety of methods including, but not limited to mechanical damage, chemical treatment, or enzymatic digestion. The polyhydroxyalkanoate polymer preferably has a molecular weight distribution of between about 1 and about 5, preferably between about 1.5 and about 4.5, more preferably between about 2 and about 4, and most preferably about 2.1 or about 2.5. The polyhydroxyalkanoate polymer may be a homopolymer or a copolymer. The polyhydroxyalkanoate polymer may generally be any polyhydroxyalkanoate polymer compatible with the inventive processes, and more preferably a polymer of 3-hydroxybutyrate, 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, 3-hydroxydodecanoate, 3-hydroxytetradecanoate, 3-hydroxyhexadecanoate, 3-hydroxyoctadecanoate, 3-hydroxyeicosanoate, 3-hydroxydocosanoate, or copolymers thereof. In more preferred embodiments, the homopolymer is poly(3-hydroxybutyrate) or poly(4-hydroxybutyrate), and the to copolymer is poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3-hydroxybutyrate-co-4-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyhexanoate), poly(4-hydroxybutyrate-co-3-hydroxyhexanoate), or poly(3-hydroxybutyrate-co-4-hydroxybutyrate-co-3-hydroxyhexanoate). The plant in which the polyhydroxyalkanoate polymer is biosynthesized is generally any plant suitable for the biosynthesis of polyhydroxyalkanoate polymers, and more preferably is tobacco, wheat, is potato, Arabidopsis, corn, soybean, canola, oil seed rape, sunflower, flax, peanut, sugarcane, swtichgrass, or alfalfa. The number average molecular weight of the polyhydroxyalkanoate polymer is preferably greater than about 100,000, more preferably greater than about 300,000, and most preferably greater than about 500,000.
The invention further provides methods for the preparation of polyhydroxyalkanoate polymers having a single mode molecular weight distribution. A preferred embodiment comprises the steps of (a) inserting into a plant cell nucleic acid molecules comprising a polyhydroxyalkanoate synthesis pathway, preferably comprising a .beta.-ketoacyl reductase gene, a .beta.-ketothiolase gene, and a polyhydroxyalkanoate synthase gene; (b) isolating a transformed plant cell; (c) regenerating the transformed plant cell to form a transformed plant; (d) selecting a transformed plant which produces a polyhydroxyalkanoate polymer having a single mode molecular weight distribution; and (e) isolating the polyhydroxyalkanoate polymer. The .beta.-ketoacyl reductase, .beta.-ketothiolase, and polyhydroxyalkanoate synthase genes may generally be of any source suitable for participation in the inventive processes, more preferably the genes are Alcaligenes eutrophus, Alcaligenes faecalis, Aphanothece sp., Azotobacter vinelandii, Bacillus cereus, Bacillus megaterium, Beijerinkia indica, Derxia gummosa, Methylobacterium sp., Microcoleus sp., Nocardia corallina, Pseudomonas cepacia, Pseudomonas extorquens, Pseudomonas oleovorans, Rhodobacter sphaeroides, Rhodobacter capsulatus, Rhodospirillum rubrum, or Thiocapsa pfennigii genes, and most preferably are Alcaligenes eutrophus genes. The polyhydroxyalkanoate polymer preferably has a molecular weight distribution of between about 1 and about 5, preferably between about 1.5 and about 4.5, more preferably between about 2 and about 4, and most preferably about 2.1 or about 2.5. The polyhydroxyalkanoate polymer may be a homopolymer or a copolymer. The polyhydroxyalkanoate polymer may generally be any polyhydroxyalkanoate polymer compatible with the inventive processes, and more preferably a polymer of 3-hydroxybutyrate, 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, 3-hydroxydodecanoate, 3-hydroxytetradecanoate, 3-hydroxyhexadecanoate, 3-hydroxyoctadecanoate, 3-hydroxyeicosanoate, 3-hydroxydocosanoate, or copolymers thereof. In more preferred embodiments, the homopolymer is poly(3-hydroxybutyrate) or poly(4-hydroxybutyrate), and the copolymer is poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3-hydroxybutyrate-co-4-hydroxybutyrate), poly(3-hydroxybutyrate-co-3-hydroxyhexanoate), poly(4-hydroxybutyrate-co-3-hydroxyhexanoate), or poly(3-hydroxybutyrate-co-4-hydroxybutyrate-co-3-hydroxyhexanoate). The plant in which the polyhydroxyalkanoate polymer is biosynthesized is generally any plant suitable for the biosynthesis of polyhydroxyalkanoate polymers, and more preferably is tobacco, wheat, potato, Arabidopsis, corn, soybean, canola, oil seed rape, sunflower, flax, peanut, sugarcane, swtichgrass, or alfalfa. The number average molecular weight of the polyhydroxyalkanoate polymer is preferably greater than about 100,000, more preferably greater than about 300,000, and most preferably greater than about 500,000. The transformed plant cells and plants may further comprise nucleic acid molecules having genes that allow the plant to synthesize additional polyhydroxyalkanoate precursors. In a preferred embodiment, the nucleic acid molecules further comprise succinyl-CoA:acetyl-CoA CoA transferase, succinate semialdehyde dehydrogenase, 4-hydroxybutyrate dehydrogenase, and hydroxybutyryl-CoA:acetyl-CoA CoA transferase genes. The inventive methods may be further extended by including a step of generating a homozygous daughter plant derived from the transformed plant. Conventional methods such as growing daughter plants from seeds, and cross-breeding may be used to generate such a homozygous daughter plant. The transformed plant may be analyzed to determine the copy number of the polyhydroxyalkanoate synthase gene. It is preferable that the synthase gene be present in a low copy number, more preferably less than five, and most preferably present at a single copy. Copy number may be determined by any method known to those of skill in the art, and preferably by Southern blotting.
The invention further provides methods for selecting a transformed plant cell or plant that is particularly suitable for the production of a polyhydroxyalkanoate polymer having a single mode molecular weight distribution. A preferred embodiment comprises the steps of (a) obtaining transformed plant cells or transformed plants, (b) analyzing the plant cells or plants for the presence of a polyhydroxyalkanoate synthase gene, (c) determining the copy number of the polyhydroxyalkanoate synthase gene, and (d) selecting a transformed plant cell or transformed plant having a single copy of the polyhydroxyalkanoate synthase gene. Copy number may be determined by any method known to those of skill in the art, and preferably by Southern blotting.
Further scope of the applicability of the present invention will become apparent from the detailed description and drawings provided below. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the present invention, are given by way of illustration only since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.